This invention relates to recombinant fucosyltransferases, DNA, and uses thereof.
The structurally related endothelial cell receptors E-selectin (Bevilacqua et al., Proc. Natl. Acad. Sci. USA 84:9238, 1987; Bevilacqua et al., Science 243:1160, 1989) and P-selectin (Hsu-Lin et al., J. Biol. Chem. 259:9121, 1984; Stenberg et al., J. Cell Biol. 101:880, 1985; Johnston et al., Blood 69:1401, 1987) mediate myeloid cell attachment to the vascular wall following activation of the endothelial cell by inflammatory cytokines (in the case of E-selectin; see, e.g., Bevilacqua et al. (1987), supra) or thrombin (in the case of P-selectin; see, e.g., Geng et al., Nature 343:757, 1990). Expressed both by Weibel-Palade bodies of endothelium (McEver et al., J. Clin. Invest. 84:920, 1989; Bonfanti et al., Blood 73:1109,.1989) and platelet alpha granules (Hsu-Lin et al., supra; Stenberg et al., supra), P-selectin also mediates monocyte and neutrophil binding to activated platelets (Larsen et al., Cell 59:305, 1989; Hamburger et al., Blood 75:550, 1990). The leading candidate ligands for the two receptors are the sialyl-Le.sup.x structure for E-selectin (Lowe et al., Cell 63:475, 1990; Phillips et al., Science 250:1130, 1990); Walz et al., Science 250:1132, 1990) and the same glycan in the context of a specific mucin, PSGL-1, (P-selectin glycoprotein ligand 1) for P-selectin (Sako et al., Cell 75:1179, 1993).
Genetic and biochemical data have demonstrated the existence of at least four distinct types of fucosyltransferases capable of forming the .alpha.(1,4) linkage: the Lewis enzyme (Fuc-TIII), which can transfer fucose either .alpha.(1,3) or .alpha.(1,4) to Gal.beta.4GlcNAc or Gal.beta.3GlcNAc respectively (Kukowska-Latallo et al., Genes Dev. 4:1288, 1990); at least one enzyme, Fuc-TIV, solely forming .alpha.(1,3) linkages, which cannot utilize sialylated substrates (Goelz et al., Cell 63:1349, 1989; Lowe et al., J. Biol. Chem. 266:17467, 1991); at least two enzymes, Fuc-TV (Weston et al., J. Biol. Chem. 267:4152, 1992a) and Fuc-TVI (Weston et al., J. Biol. Chem. 267:24575, 1992b) solely forming .alpha.(1,3) linkages, which can fucosylate either sialylated or nonsialylated precursors, and a recently described enzyme, Fuc-TVII, (Sasaki et al., J. Biol. Chem. 269:14730, 1994); Natsuka et al., J. Biol. Chem. 269:16789, 1994) which can fucosylate only sialylated precursors.